Background: Lately, several studies have utilized non-invasive serological markers to assess liver fibrosis and some are currently being validated as potential tools to determine liver damage. Purpose: Our aim was to investigate the diagnostic performance of AFP, autotoxin and collagen IV as non-invasive biomarkers of hepatic fibrosis. Patients and methods: 45 males and 15 females with chronic hepatitis C were enrolled in the current study. Laboratory assessment was done for all subjects in form of complete blood picture, liver function test, alpha fetoprotein (AFP), collagen IV and autotaxin. Patients were grouped according to the stage of fibrosis into F1, F2 and F3. Results: Mean serum values of AFP, autotaxin and collagen IV were elevated in all patients compared to healthy controls. Surprisingly, with increasing fibrosis stage AFP showed non-significant change while collagen IV and autotoxin showed significant increase (P less than 0.01 and P less than 0.0001, respectively). Autotaxin and collagen IV were significantly (P less than 0.01 and P less than 0.05, respectively) lower in F1 patients than those with F2-F3 but AFP level showed non-significant change. Autotaxin had the highest area under ROC curve and the highest accuracy for discrimination of F1 from F2-F3 patients and for discrimination of patients with F3 from F1-F2. Different combinations between AFP, collagen IV and autotoxin showed improvement in the accuracy. Conclusion: It was concluded that serum autotaxin may at least serve as a new clinical non-invasive alternative in patients who are not candidates for liver biopsy for diagnosis of liver damage. Autotaxin combination with collagen IV and AFP addition make them more useful.